Corona ion generating method and apparatus for the manipulation of molecules and biological cells

Abstract

The present invention provides a method and apparatus adapted to facilitate the entry of a preselected molecule into the intracellular space of a cellular sample through the use of ions generated by a corona charge source. With the present method and apparatus, molecules are manipulated within cells and in the extracellular space surrounding the cells. Manipulation enhances the permeability of cell barriers to allow the subsequent introduction of molecules of interest into the interior of a cell.

1. A method to facilitate the manipulation of molecules of a biological cellular sample, the method comprising the steps of:
 providing a biological cellular sample;  
 providing a corona charge source;  
 positioning the corona charge source proximate to the cellular sample; and  
 establishing a corona charge in proximity to the cellular sample whereby the establishment of the corona charge is effective in manipulating the molecules of the cellular sample.  
 
     
     
       2. The method of  claim 1 , further comprising the step of applying a bias potential to the cellular sample. 
     
     
       3. The method of  claim 1 , further comprising the step of introducing a preselected molecule into the extracellular space of the cellular sample. 
     
     
       4. The method of  claim 3 , further comprising the step of manipulating the preselected molecule within the extracellular space. 
     
     
       5. The method of  claim 3 , further comprising the step of inducing movement of the preselected molecule in the extracellular space such that the molecule passes through the permeable cell membrane into the interior of the cell. 
     
     
       6. The method of  claim 1 , further comprising the step of inducing movement of an intracellular molecule into the extracellular space of the cellular sample. 
     
     
       7. The method of  claim 1 , further comprising the step of moving the corona charge source in a predetermined direction relative to the cellular sample. 
     
     
       8. The method of  claim 1 , further comprising the step of moving the cellular sample in a predetermined direction relative to the corona charge source. 
     
     
       9. A method of fusing two or more biological cells, comprising the steps of:
 providing a biological cellular sample;  
 providing a corona charge source;  
 forcing contact between two or more cells of the cellular sample; and  
 positioning the corona charge source proximate to the cellular sample, whereby fusion of the two or more cells is effected.  
 
     
     
       10. The method of  claim 9 , further comprising the step of applying a bias potential to the cellular sample. 
     
     
       11. A method to facilitate the entry of a preselected molecule into the intracellular space of a biological cellular sample, the method comprising the steps of:
 providing a biological cellular sample, the sample comprising cells and extracellular space;  
 providing a corona charge source;  
 establishing a corona charge in proximity to the cellular sample to increase the permeability of the cell membrane; and  
 inducing movement of a preselected molecule in the extracellular space such that the molecule passes through the permeable cell membrane into the intracellular space of a cell of the cellular sample.  
 
     
     
       12. The method of  claim 11 , further comprising the step of applying a bias potential to the cellular sample. 
     
     
       13. The method of  claim 11  further comprising the step of introducing the preselected molecule into the extracellular space of the cellular sample. 
     
     
       14. An apparatus to facilitate the manipulation of molecules of a biological cellular sample, the apparatus comprising:
 a support member; and  
 at least one biological molecule manipulating corona charge emitting device extending away from the support member, the biological molecule manipulating corona charge emitting device generating a corona charge in proximity to the cellular sample for the effective manipulation of the molecules of the biological cellular sample.  
 
     
     
       15. The apparatus of  claim 14  wherein the corona charge emitting device is affixed to the support member. 
     
     
       16. The apparatus of  claim 14  wherein the corona charge emitting device defines the support member. 
     
     
       17. The apparatus of  claim 14  further comprising a spacer circumferentially encompassing the corona charge emitting device, the spacer defining the proximity of the corona charge emitting device relative to the cellular sample. 
     
     
       18. The apparatus of  claim 17  wherein the spacer is substantially transparent. 
     
     
       19. The apparatus of  claim 14  further comprising a restrictor circumferentially encompassing the corona charge emitting device, the restrictor defining a restricted area of the cellular sample in which the corona charge is generated. 
     
     
       20. The apparatus of  claim 19  wherein the restricted area is a predetermined geometric configuration. 
     
     
       21. The apparatus of  claim 19 , wherein the restrictor is substantially transparent. 
     
     
       22. The apparatus of  claim 19 , whereby contact between the restrictor and the cellular sample establishes a substantially airtight cavity. 
     
     
       23. The apparatus of  claim 22 , further comprising an atmospheric controller in communication with the airtight cavity, the atmospheric controller adapted to regulate the air within the cavity. 
     
     
       24. The apparatus of  claim 14 , further comprising a portal surrounded by the support member, the portal positioned to accommodate a device adapted for the introduction of a molecule of interest into the cellular sample. 
     
     
       25. The apparatus of  claim 24 , wherein the device adapted for the introduction of a molecule of interest is selected from the group consisting of, a hypodermic needle, or a jet injection apparatus. 
     
     
       26. The apparatus of  claim 14 , further comprising a reservoir positioned between the corona charge emitter and the cellular sample, the reservoir containing a molecule of interest. 
     
     
       27. The apparatus of  claim 14 , further comprising a cellular sample holding apparatus to contain the cellular sample. 
     
     
       28. The apparatus of  claim 27 , further comprising means for moving the corona charge emitting device in a predetermined direction relative to the cellular sample. 
     
     
       29. The apparatus of  claim 27 , further comprising means for moving the cellular sample in a predetermined direction relative to the corona charge emitting device.